biotek elx808 absorbance microplate reader -...
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BioTek ELx808
Absorbance Microplate Reader
Bergen County Technical Schools Biotechnology Lab
Room 224A
BioTek ELx808 Absorbance Microplate Reader Information
The BioTek ELx808 Absorbance Microplate Reader is a multi-channel
absorbance microplate reader that can be used for research and development and
diagnostic use. The instrument essentially functions as a spectrophotometer, which
measures intensity as a function of the color, or more specifically, the wavelength of
light. The instrument has applications that would provide useful for multiple disciplines,
from Biology, to Chemistry and Nanotechnology.
Unlike other microplate readers, the ELx808 does not support a wide range of
labware. The ELx808 has been designed to only read 96 well plates.
The KC junior software that is installed with the instrument controls all of the
measurements, actions, and protocols performed by the ELx808. The software supports
detection of only absorbance, with applications that have been designed for use in
areas such as Agriculture, Biotech Research, Cell Biology, Cell Counting, Cell
Proliferation, ELISA, the Environment, and Neuroscience.
The software allows the user to establish unique protocols to analyze different
sample plates. Samples are analyzed in the instrument, and the results stored on the
computer’s hard drive. The data from the experiments are able to be viewed and
analyzed in the KC junior program, as well as in Excel by exporting the data.
1. Turn on the BioTek ELx808 Absorbance Microplate Reader by
BioTek ELx808 Absorbance Microplate Reader Quick Start Guide
flipping the Power Switch, located on lower right side of the
instrument.
a. Allow the instrument to perform its start-up procedure before proceeding.
The screen on the instrument will show “Powerup Sequence V1.02
Initializing” while the instrument is starting up.
b. When the instrument stops making noise, it is ready to use. The screen on
the instrument will show “Ready” when it is ready to use.
2. On the desktop, click the KC junior Icon to start the software.
Note: The software will not work correctly if started before
the instrument’s screen displays “Ready”.
Note: The user must be logged in under donleo on the computer in order
for the software to work correctly with the instrument. Conditions have
been optimized in this user setting. If the computer is not logged in as
donleo, ask the instructor for assistance.
a. Before being able to run anything in the program, a screen will pop-up
where the user needs to click OK. This will allow the instrument to initialize
and communicate with the software.
1. Once the program starts, the user will have options such as
Setting up a Protocol
Read Plate or Open Results, and New, Open, or Modify a
Protocol on the Main Menu Screen.
2. Click New Protocol to create a new protocol.
3. On the Protocol Definition screen that appears, select the
General Information Tab.
a. Enter the Protocol Name in the area labeled.
i. The protocol name should include the user’s name, the sampleor
experiment, and the data. (Ex StudentX_ELISA_1/1/09).
b. Enter a Protocol Description.
i. Information about the type of samples being run, wavelength’s
being used, and other general information can be included in this
area.
c. The remaining portions of this tab do not need to be changed.
Note: Do not click OK at the bottom of the screen until all tabs have been
filled in. Only at the end of making the protocol should the OK button be
pressed.
4. Select the Read Methods Tab.
a. First select the Read Method Type that will be used for the plate.
i. Endpoint is the type most often used. This method scans the wells
at the wavelength selected.
ii. Kinetic is used for plates that need to be read multiple times over a
set period of time.
iii. MultiWavelength is used for plates that need to be scanned at
more than one wavelength.
b. Select the Primary Wavelength from the Dropdown Menu.
i. For this instrument, the plates can be read at 405nm, 450nm,
490nm, or 630nm.
c. A Reference Wavelength does not need to be set for the plate.
d. The plate can be delayed before the read
takes place by entering a time in the box
next to Delay Before Read (sec).
e. The instrument can be used to shake the plate prior to reading by
selecting a Shake Method from the Dropdown Menu.
i. The Intensity of the shake can be changed in the Dropdown Menu.
ii. The amount of time in seconds to shake the plate can be entered in
the box Duration.
f. The temperature cannot be set for this instrument, so the area Incubation
and Temperature cannot be selected or edited.
g. Once all of the read conditions for the plate are entered, click the Validate Read Method Button to make sure
that the protocol can be run correctly.
i. If the read method is ok, a pop-up will tell the user that the read
method is valid.
ii. If the read method is not ok, the pop-up will instruct the user on
what to change in the protocol.
5. Select the Template Tab.
a. In the template tab, the contents of each well in the plate can be assigned.
b. Make sure that the Well ID’s Button is selected.
c. First select Standards from the Dropdown Menu
under Well Type Selections.
d. If there are replicates of the standards in the plate,
select the number of replicates, and if they go
down or across in Replicates / Direction.
e. The Sample Number can be changed if the user does
not want the number to start at one.
f. On the template, click on a box to apply a standard or replicates of a
standard to that box.
g. Repeat steps c – f for Samples, Blank, Controls, and Empty wells in the
plate. Repeat until the entire plate template is established.
h. Click the Concentration Button to set the
concentration of the standards.
i. Click on a standard and enter its concentration. Repeat for each
standard.
ii. Selecting the Delta Button will increase the next
standard by the value entered.
iii. Selecting the Factor Button will increase the next
standard by a factor of the value entered.
iv. Click OK.
i. Click the Dilutions Button at the top of the screen if
there are dilutions of a sample in the plate. This can
only be applied to wells with a sample or control in it.
i. Select the sample and enter the dilution factor.
j. Click the Transformation Button at the top of the
screen if formulas need to be applied to the results of
samples in certain wells.
i. Select the sample and enter the formula.
k. Click the Sample IDs Button at the top of the screen to
give a name to samples selected in the plate.
l. Make sure a template has been established for the entire plate.
6. Click the Curve Tab to set up the parameters of the standard
curve that can be developed from the run.
a. Select a Curve Method from the Dropdown Menu.
i. Linear is the type of graph used for most
runs. Other graph options include Quadratic,
Cubic, 2 Parameter, 4 Parameter, Point-to-
Point, or Cubic Spine.
b. Enter a Graph Report Name.
i. The name should say “Standard Curve of …” whatever the
standard used in the plate is.
c. For the X and the Y axis, select if the scale
should be Linear or Logarithmic.
d. Enter a Label for each axis if the user wants a unique name for each.
7. The sections Cutoffs and Reports do not need to be changed.
8. Click OK once the entire protocol is completed.
9. Before running the protocol, it must be saved.
a. Click the Save Protocol Icon on the right side of the Toolbar (the
save icon on the left side is for experiments).
b. Name the protocol in the provided directory with the user name, type of
sample or experiment, and the date.
i. Ex. StudentX_ELISA_1/1/09 Protocol
Once the protocol is saved, it can be modified or opened at a
later time using the options on the first menu page. If the protocol
will not be run until a later data or time, it must be opened prior to
starting a new run.
1. Make sure that an established protocol has been opened in the program.
Running a Protocol on a Sample Plate
2. Click the Read Plate Button on the Main Menu Screen.
3. A screen will pop-up. Enter a Results ID in the box.
a. The results ID should include the student’s name, sample, and date.
4. A Plate Description does not need to be entered, but can
be with information about the plate being read.
5. Click Read Plate once the information has been entered.
6. Place the microplate in the carrier.
7. On the pop-up screen, re-enter the result ID that was already given in the area
labeled Plate ID.
8. Make sure the plate is on the carrier, and click OK. The instrument will read the
absorbance of the samples in the plate.
1. A screen will pop-up after the run is completed. This contains all of the data from
the run.
Analyzing the Results of the Plate
a. Click the Raw Data Tab. This will provide the user with
each well’s actual reading from the instrument.
b. Click the Blanked Data Tab. This subtracts the average
value from the wells labeled blank in the template from
all other wells in the plate.
c. Click the Concentrations Tab. This shows the concentration of each of
the samples in the plate based on the absorbance
reading and the values of the standards on the plate.
d. Click the Template Tab to see what was in each of the
wells in the plate.
2. Click the Statistics Button. This lists all of the data in
list form. It also groups replicates of samples and gives
the average value of the replicates.
3. Click the Standard Curve Button.
a. This button plots all of the standards from the plate.
b. An Equation for the Line is given
on the graph. Sample
concentrations can be determined
by entering the absorbance readings into the equation.
c. The parameters set in the protocol for the curve
can be adjusted by clicking the Curve Params… Button, and the standard concentrations can be
adjusted by clicking the Edit Stds… Button.
d. Click Store the Curve to save the curve. Make
sure to name it with the Student’s name, sample or
experiment, and the date.
e. The graph and information can be printed from this
screen by clicking the Print / Copy Button.
4. To export the information from the run to Excel, click Results Export Data on
the Toolbar.
a. Select the different types of data to export to Excel by
selecting each and clicking the Add Button.
b. Click the Export Button to send the data to
Excel.
c. In Excel, the data will show up in a format that matches the wells of the
microplate, and will be labeled with what type of information is contained
in each set.
Note: Results from runs can be accessed anytime after
they are saved by clicking the Open Results Button
on the Main Menu Screen.
1. When finished with the experiment and data, click the Save
Saving Data and Shutting Down the Instrument
Experiment Icon on the left side of the Toolbar to save the results.
a. Enter a name for the experiment in the directory that appears. Include the
student’s name, sample or experiment type, and the date.
2. Shut down the KC junior program by clicking the (X) Button in the
top right corner of the screen.
3. Make sure that the microplate was removed from the
instrument, and turn the instrument off by pressing the
Power Switch on the lower right side of the instrument.
4. Make sure the station is neat before leaving the instrument!
1. Samples should be handled according to good laboratory procedures and methods in order to prevent accidents.
BioTek ELx808 Absorbance Microplate Reader Safety Sheet
2. Dispose of all waste solutions according to waste disposal procedures.
3. Do not remove any panels or cords from the instrument to avoid electrical shock.
4. The instrument has moving parts:
a. Do not attempt to exchange microplates while the instrument is operating.
b. Keep the work area around the instrument clear to avoid obstruction of the moving parts.
c. When the microplate carriage is open, only place a microplate in the carriage to avoid breaking the instrument.
d. Be aware of clothing and body parts when instrument doors are opening and closing.
5. Cleaning the workstation around the instrument is necessary. Never attempt to clean any internal spaces of the instrument.
6. Measurement values may be affected by extraneous particles (such as dust) in the microplate wells. A clean work area is necessary for an accurate reading.
7. Food and drinks should not be placed on or near the instrument.
8. If any liquid should fall near the instrument, do not operate the instrument. Fluid seepage into internal components creates a potential shock hazard, and can cause the instrument to not work properly.
9. Running unauthorized programs or changing preferences on the instrument’s computer is not allowed. Laptops are provided for personal use and internet access. Do not plug memory sticks in the instrument’s computer unless instructed to do so by the instructor.
BioTek ELx808 Absorbance Microplate Reader Instrument Information
ELx808 Absorbance Microplate Reader Overview:
1. Instrument Information Screen
2. Instrument Information Entry Keys
3. Power Switch
4. Microplate Carrier Door
5. Microplate Carrier
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